Details

Autor(en) / Beteiligte

• George, Kevin W.
• Thompson, Mitchell G.
• Kim, Joonhoon
• Baidoo, Edward E.K.
• Wang, George
• Benites, Veronica Teixeira
• Petzold, Christopher J.
• Chan, Leanne Jade G.
• Yilmaz, Suzan
• Turhanen, Petri
• Adams, Paul D.
• Keasling, Jay D.
• Lee, Taek Soon

Titel

Integrated analysis of isopentenyl pyrophosphate (IPP) toxicity in isoprenoid-producing Escherichia coli

Ist Teil von

• Metabolic engineering, 2018-05, Vol.47, p.60-72

Ort / Verlag

Belgium: Elsevier Inc

Erscheinungsjahr

2018

Link zum Volltext

Quelle

Elsevier ScienceDirect Journals Complete

Beschreibungen/Notizen

• Isopentenyl pyrophosphate (IPP) toxicity presents a challenge in engineered microbial systems since its formation is unavoidable in terpene biosynthesis. In this work, we develop an experimental platform to study IPP toxicity in isoprenol-producing Escherichia coli. We first characterize the physiological response to IPP accumulation, demonstrating that elevated IPP levels are linked to growth inhibition, reduced cell viability, and plasmid instability. We show that IPP toxicity selects for pathway “breakage”, using proteomics to identify a reduction in phosphomevalonate kinase (PMK) as a probable recovery mechanism. Next, using multi-omics data, we demonstrate that endogenous E. coli metabolism is globally impacted by IPP accumulation, which slows nutrient uptake, decreases ATP levels, and perturbs nucleotide metabolism. We also observe the extracellular accumulation of IPP and present preliminary evidence that IPP can be transported by E. coli, findings that might be broadly relevant for the study of isoprenoid biosynthesis. Finally, we discover that IPP accumulation leads to the formation of ApppI, a nucleotide analog of IPP that may contribute to observed toxicity phenotypes. This comprehensive assessment of IPP stress suggests potential strategies for the alleviation of prenyl diphosphate toxicity and highlights possible engineering targets for improved IPP flux and high titer isoprenoid production. •Three strains at various IPP accumulation levels were comparatively analyzed.•IPP accumulation was linked to decreased cell viability and nutrient uptake.•IPP-induced pathway “breakage” was characterized using multi-omics data.•Nucleotide metabolism and ATP synthesis were perturbed by IPP accumulation.•A nucleotide analog of IPP was identified as a possible contributor to toxicity.